MEASURING c-Abl ACTIVITY IN CML USING A HIGH-AFFINITY NANOSENSOR

描述（由申请人提供）：监测和测量肿瘤发生和癌症中蛋白激酶活性的能力对转化过程具有指示性和关键性，因此代表了一种有吸引力的诊断策略，具有数十亿美元的市场机会。在这项提议中，我们的目标是开发一种新型的，高亲和力的荧光纳米传感器和均匀的检测系统，用于监测c-Abl的真实肽底物磷酸化，c-Abl是一种重要的激酶，参与慢性粒细胞白血病（CML）的病因学。在提案的这一阶段部署的技术将直接转化为CML诊断敏感平台的开发。在具体目标1中，我们将合成高亲和纳米传感器。在特定目标2中，我们将使用该试剂开发一种敏感的一步检测方法。具体目标3将详细说明这一点，并测试纳米传感器在测量cml阳性细胞裂解物中c-Abl活性方面的有效性。考虑到大约有400种疾病，如癌症，与蛋白激酶有关，开发一个敏感的纳米传感器家族，如本拨款申请中提出的1，将有助于这些疾病的敏感和有选择性的诊断，因此变得至关重要，并将在我们医疗保健的各个方面，从药物发现到患者健康和护理点诊断，发现巨大的效用。

DESCRIPTION (provided by applicant): The ability to monitor and measure protein kinase activity in tumorigenesis and cancer can be indicative of and critical to the transformation process and therefore represents an attractive diagnostic strategy, with a multi-billion dollar market opportunity. In this proposal, we aim to develop a novel, high affinity fluorescent nanosensor and homogeneous assay system for monitoring the phosphorylation of a bona fide peptide substrate by c-Abl, an important kinase involved in the etiology of chronic myelogenous leukemia (CML), using FRET. The technology deployed during this phase of the proposal will directly translate into the development of a sensitive platform for the diagnosis of CML. In specific aim 1, we will synthesize the high-affinity nanosensor. In specific aim 2, we will develop a sensitive 1-step assay using this reagent. Specific aim 3 will elaborate on this and test the effectiveness of the nanosensor in measuring c-Abl activity in CML-positive cell lysates. Given the fact that ~400 disorders such as cancer have been associated with protein kinases, the development of a family of sensitive nanosensors such as the 1 proposed in this grant application will facilitate the diagnosis of these diseases sensitively and selectively and therefore becomes of paramount importance and will find immense utility in all the various facets of our healthcare, from drug-discovery to patient health and point-of-care diagnostics.